| 质粒名称: | pCS-H2B-EGFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 53744 |
| 存储实验室: | Sean Megason |
|
载体骨架基本信息 |
|
| 载体名称: | pCS2+ |
| 载体出品公司: | RZPD |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 4095 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | histone H2B,histone cluster 2, H2be |
| 物种来源: | H. sapiens (human) |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | simian CMV IE94 |
| 标签/融合蛋白: | EGFP（插入件上的C端子） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | SnaB1（克隆过程中销毁） |
| 3' 克隆位点: | SnaB1（克隆过程中销毁） |
| 5' 测序引物 | SP6 |
| 3' 测序引物 | EBV-rev |
|
文献引用方法 |
|
| 材料和方法部分: | pCS-H2B-EGFP was a gift from Sean Megason (Addgene plasmid # 53744 ; http://n2t.net/addgene:53744 ; RRID:Addgene_53744) |
| 参考文献部分: | In toto imaging of embryogenesis with confocal time-lapse microscopy. Megason SG. Methods Mol Biol. 2009;546:317-32. doi: 10.1007/978-1-60327-977-2_19. 10.1007/978-1-60327-977-2_19 PubMed 19378112 |
买家导航
pCS-H2B-EGFP载体质粒基本信息
pCS-H2B-EGFP质粒图谱载体图谱和pCS-H2B-EGFP载体序列质粒序列多克隆位点信息
pCS-H2B-EGFP质粒载体简介
pCS-H2B-EGFP质粒序列载体序列
LOCUS pCS-H2B-EGFP 5260 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Contains human Histone H2B fused to EGFP in the pCS2+ expression
vector.
ACCESSION .
VERSION .
KEYWORDS pCS-H2B-EGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5260)
AUTHORS Megason SG
TITLE In toto imaging of embryogenesis with confocal time-lapse
microscopy.
JOURNAL Methods Mol Biol. 2009;546:317-32. doi:
10.1007/978-1-60327-977-2_19.
PUBMED 19378112
REFERENCE 2 (bases 1 to 5260)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5260
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(45..65)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(45..63)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(84..100)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(84..100)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(97..119)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 108..124
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(132..162)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 177..198
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(315..332)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(486..1074)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(566..585)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(1245..2105)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 1868..1887
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(2106..2210)
/gene="bla"
/label=AmpR promoter
rep_origin complement(2236..2691)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2373..2394)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2585..2604
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2765..3749
/label=CMV IE94 promoter
/note="enhancer/promoter region of simian cytomegalovirus
major immediate early transcription unit IE94"
promoter 3784..3802
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 3784..3801
/label=SP6
/note="SP6 promoter, forward primer"
promoter complement(3871..3888)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 3907..4284
/codon_start=1
/gene="HIST1H2BJ"
/product="human histone H2B"
/label=H2B
/translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
GELAKHAVSEGTKAITKYTSAK"
regulatory 4297..4306
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 4303..5022
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(4348..4369)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(4609..4628)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 4956..4977
/label=EGFP-C
/note="EGFP, forward primer"
promoter complement(5036..5053)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
polyA_signal 5058..5192
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5082..5101)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 5136..5155
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
ORIGIN
1 ggccgcggcg ccaatgcatt gggcccggta cccagctttt gttcccttta gtgagggtta
61 attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
121 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
181 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
241 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
301 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
361 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
421 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
481 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
541 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
601 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
661 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
721 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
781 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
841 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
901 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
961 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
1021 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
1081 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
1141 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
1201 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
1261 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
1321 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
1381 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
1441 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
1501 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
1561 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
1621 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
1681 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
1741 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
1801 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
1861 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
1921 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
1981 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
2041 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
2101 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
2161 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
2221 cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
2281 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
2341 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
2401 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
2461 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
2521 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
2581 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
2641 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc
2701 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
2761 agtcgaccat agccaattca atatggcgta tatggactca tgccaattca atatggtgga
2821 tctggacctg tgccaattca atatggcgta tatggactcg tgccaattca atatggtgga
2881 tctggacccc agccaattca atatggcgga cttggcacca tgccaattca atatggcgga
2941 cttggcactg tgccaactgg ggaggggtct acttggcacg gtgccaagtt tgaggagggg
3001 tcttggccct gtgccaagtc cgccatattg aattggcatg gtgccaataa tggcggccat
3061 attggctata tgccaggatc aatatatagg caatatccaa tatggcccta tgccaatatg
3121 gctattggcc aggttcaata ctatgtattg gccctatgcc atatagtatt ccatatatgg
3181 gttttcctat tgacgtagat agcccctccc aatgggcggt cccatatacc atatatgggg
3241 cttcctaata ccgcccatag ccactccccc attgacgtca atggtctcta tatatggtct
3301 ttcctattga cgtcatatgg gcggtcctat tgacgtatat ggcgcctccc ccattgacgt
3361 caattacggt aaatggcccg cctggctcaa tgcccattga cgtcaatagg accacccacc
3421 attgacgtca atgggatggc tcattgccca ttcatatccg ttctcacgcc ccctattgac
3481 gtcaatgacg gtaaatggcc cacttggcag tacatcaata tctattaata gtaacttggc
3541 aagtacatta ctattggaag gacgccaggg tacattggca gtactcccat tgacgtcaat
3601 ggcggtaaat ggcccgcgat ggctgccaag tacatcccca ttgacgtcaa tggggagggg
3661 caatgacgca aatgggcgtt ccattgacgt aaatgggcgg taggcgtgcc taatgggagg
3721 tctatataag caatgctcgt ttagggaacc gccattctgc ctggggacgt cggagcaagc
3781 ttgatttagg tgacactata gaatacaagc tacttgttct ttttgcagga tcccatcgat
3841 tcgaattcaa ggcctctcga gcctctagaa ctatagtgag tcgtattact cgacggtacc
3901 gccaccatgc cagagccagc gaagtctgct cccgccccga aaaagggctc caagaaggcg
3961 gtgactaagg cgcagaagaa aggcggcaag aagcgcaagc gcagccgcaa ggagagctat
4021 tccatctatg tgtacaaggt tctgaagcag gtccaccctg acaccggcat ttcgtccaag
4081 gccatgggca tcatgaattc gtttgtgaac gacattttcg agcgcatcgc aggtgaggct
4141 tcccgcctgg cgcattacaa caagcgctcg accatcacct ccagggagat ccagacggcc
4201 gtgcgcctgc tgctgcctgg ggagttggcc aagcacgccg tgtccgaggg tactaaggcc
4261 atcaccaagt acaccagcgc taaggatcca ccggtcgcca ccatggtgag caagggcgag
4321 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
4381 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
4441 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
4501 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
4561 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
4621 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
4681 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
4741 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
4801 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
4861 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
4921 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
4981 gccgccggga tcactctcgg catggacgag ctgtacaagt aaagcggccc tagaactata
5041 gtgagtcgta ttacgtagat ccagacatga taagatacat tgatgagttt ggacaaacca
5101 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
5161 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
5221 ttcaggttca gggggaggtg tgggaggttt tttaattcgc
//
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